Biopharmaceutics and Formulation Development Laboratory (B-ForDev Lab)

Macroarea: Pharmaceutical Technology

ERC Sectors: LS7_4 Regenerative medicine; LS1_1 Macromolecular complexes including interactions involving nucleic acids, proteins, lipids and carbohydrates; PE5_7 Biomaterials synthesis; PE5_6 New materials: oxides, alloys, composite, organic-inorganic hybrid, nanoparticles

Lab Manager: Silvia Rossi (PO), Giuseppina Sandri (PA), Barbara Vigani (RTDb), Marco Ruggeri (RTDa)

Junior Staff: Caterina Valentino (Assegnista), Eleonora Bianchi (Dottoranda), Cristian Nomicisio (Dottorando), Daiana Ianev (Dottoranda), Simone Marsani (Dottorando), Marta Pollini (Dottoranda), Amedeo Ungolo (Dottorando), Gaia Zucca (Dottoranda)

1. Design and development of scaffolds for skin tissue repair.

Aim of this research is to design and develop flexible therapeutic platforms to enhance wound healing and to prevent or treat infection in chronic wounds. Scaffolds are prepared by means of easy scalable processes, as electrospinning, centrifugal spinning, and spray-drying; porous scaffolds produced by freeze-drying are also under investigation. These systems are designed to have a three-dimensional structure capable of homing cells and of enhancing their proliferation and extracellular matrix production to restore the native tissue. Green manufacturing processes are under development to avoid organic solvents and waste. Moreover, part of the activities is devoted to the study of innovative materials to obtain scaffolds characterized by suitable mechanical properties and complete in vivo degradation. Functional materials with advanced properties for regenerative medicine are used: in particular, polysaccharides, such as chitosan, gums, alginate, maltodextrin, dextran, and carrageenan are employed as biomaterials to enhance healing process thanks to their biocompatibility, low toxicity, and bioactivity. The biological augmentation due to biomodulators, as growth factors, is also used as additional strategy along with the loading of antimicrobial agents (among this also inorganics) are considered to enhance system effectiveness.

2. Design and development of therapeutic platforms for the treatment of nervous tissue injuries.

The aim of the research is the development of innovative therapeutic systems, endowed with both neuroprotective and neuroregenerative potential, to be applied at the site of nervous tissue injury. Such systems, thanks to their fibrous and/or multi-channel structure, mimics the architecture of the extracellular matrix, providing mechanical and trophic support to nerve cells and, thus, promoting the recovery and reorganization of neuronal connectivity. Structures characterized by porosity and channels of different dimensions are developed depending on the lesion size and the nervous tissue (central or peripheral) to be treated. Moreover, the developed therapeutic platforms can be used as delivery systems of drug candidates or biological molecules capable of reducing glutamatergic excitotoxicity, preventing oxidative damage and modulating the overexpression of pro-inflammatory cytokines at the site of injury.

3. Design and development of engineered tendon substitutes.

The research focuses on the development of biopolymers-based scaffolds, both natural and synthetic, which have the ability to mimic the structural, biomechanical, and biochemical functions of the extracellular matrix, consequently mimicking the native tissues. The scaffolds are produced by means of the electrospinning, a simple, versatile, flexible, and cost‐effective method to spin polymeric materials by means of a high voltage electric field to generate ultra‐thin fibers with diameters in the nanometric or micrometric range. In particular, electrospinning allows the generation of nanofibrous porous scaffolds suitable for the tendon tissue regeneration. Prototypes obtained using freeze drying are also under development. Moreover, the polymers are combined with various organic and inorganic materials in order to increase the systems mechanical properties and biocompatibility, enhancing the cell adhesion, proliferation and differentiation, and, consequently, the tissue healing potential.

4. Development of bioadhesive and/or in situ gelling systems.

The aim of the research is the development and characterization of bioadhesive and in situ gelling systems for the local treatment of mucosal inflammation and/or infections. Depending on both the mucosa to be treated (buccal, esophageal, vaginal, intestinal) and the pathological condition to solve, the formulations are designed to be used as delivery systems of antimicrobial agents, anti-inflammatory compounds and/or probiotics.

5. Design and development of composite systems in the treatment of glioblastoma multiforme.

The aim of the research is the design of composite systems to be locally applied in the tumor resection site. These systems are composed of lipid nanoparticles, loaded with lipophilic anti-proliferative drug candidates and embedded into a polymeric matrix (electrospun fibers or in situ-gelling and mucoadhesive formulations) of easy application. The gradual degradation/dissolution of the polymeric matrix in the biologic fluids should ensure a controlled release of the nanosystems, which, thanks to their size and surface properties, should be able to establish an intimate contact with the remaining cancer cells.

• Bianchi E., Ruggeri M., Vigani B., Del Favero E., Ricci C., Boselli C., Icaro Cornaglia A,, Viseras C., Rossi S., Sandri G. Cerium Oxide and Chondroitin Sulfate Doped Polyurethane Scaffold to Bridge Tendons. ACS Applied Material & Interfaces 2023; 15: 26510–26524. https://doi.org/10.1021/acsami.3c06144
• Valentino C., Vigani B., Zucca G., Ruggeri M., Marrubini G., Boselli C., Icaro Cornaglia A., Sandri G., Rossi S. Design of Novel Mechanically Resistant and Biodegradable Multichannel Platforms for the Treatment of Peripheral Nerve Injuries. Biomacromolecules 2023; 24: 1731-1743. https://doi.org/10.1021/acs.biomac.2c01498
• Ruggeri M., Vigani B., Boselli C., Icaro Cornaglia A., Colombo D., Sànchez-Espejo R., Del Favero E., Mandras N., Roana J., Cavallo L., Cantù L., Viseras C., Rossi S., Sandri G. Smart nano-in-microparticles to tackle bacterial infections in skin tissue engineering. Materials Today Bio 2022; 16: 100418. https://doi.org/10.1016/j.mtbio.2022.100418
• Vigani B., Valentino C., Sandri G. Caramella C.M., Ferrari F., Rossi S. Spermidine crosslinked gellan gum-based “hydrogel nanofibers” as potential tool for the treatment of nervous tissue injuries: a formulation study. International Journal of Nanomedicine 2022; 17: 3421-3439. https://doi.org/10.2147/IJN.S368960
• Ruggeri M., Bianchi E., Rossi S., Boselli C., Icaro Cornaglia A., Malavasi L., Carzino R., Suarato G., Sánchez-Espejo R., Athanassiou A., Viseras C., Ferrari F., Sandri G. Maltodextrin-amino acids electrospun scaffolds cross-linked with Maillard-type reaction for skin tissue engineering. Materials Science and Engineering 2021; Article number 112593. https://doi.org/10.1016/j.msec.2021.112593
• Vigani B., Valentino C., Sandri G., Listro R., Fagiani F., Collina S., Lanni C., Bonferoni M.C., Caramella C.M., Rossi S., Ferrari F. A Composite Nanosystem as a Potential Tool for the Local Treatment of Glioblastoma: Chitosan-Coated Solid Lipid Nanoparticles Embedded in Electrospun Nanofibers. Polymers (Basel) 2021; 13(9):1371. https://doi.org/10.3390/polym13091371
• Vigani B., Valentino C., Cavalloro V., Catenacci L., Sorrenti M., Sandri G., Bonferoni M.C., Bozzi C., Collina S., Rossi S., Ferrari F. Gellan-Based Composite System as a Potential Tool for the Treatment of Nervous Tissue Injuries: Cross-Linked Electrospun Nanofibers Embedded in a RC-33-Loaded Freeze-Dried Matrix. Pharmaceutics 2021; 13(2): 164. https://doi.org/10.3390/pharmaceutics13020164
• Sandri G., Faccendini A., Longo M., Ruggeri M., Rossi S., Bonferoni,M.C., Miele D., Prina-Mello A., Aguzzi C., Viseras C., Ferrari F. Halloysite- and Montmorillonite-Loaded Scaffolds as Enhancers of Chronic Wound Healing. Pharmaceutics 2020; 12, 179. https://doi.org/10.3390/pharmaceutics12020179
• Vigani B., Rossi S., Sandri G., Bonferoni M.C., Rui M., Collina S., Fagiani F., Lanni C., Ferrari F. Dual-Functioning Scaffolds for the Treatment of Spinal Cord Injury: Alginate Nanofibers Loaded with the Sigma 1 Receptor (S1R) Agonist RC-33 in Chitosan Films. Marine Drugs 2019; 18(1): E21. https://doi.org/10.3390/md18010021
• Sandri G., Rossi S., Bonferoni M.C., Miele D., Faccendini A., Del Favero E., Di Cola E., Icaro Cornaglia A., Boselli C., Luxbacher T., Malavasi L., Cantù L., Ferrari F. Chitosan/glycosaminoglycan scaffolds for skin reparation. Carbohydrate Polymers 2019; 220, 219-227. https://doi.org/10.1016/j.carbpol.2019.05.069

National collaborations

• Dr. A. Athanassiou - Istituto Italiano di Tecnologia
• Prof. L. Cantù e E. Del Favero – Università degli Studi di Milano Statale
• Prof. N. Mandras – Università degli Studi di Torino
• Prof. S. Riela – Università degli Studi di Palermo
• Prof. B Onida e F. Bosco – Politecnico di Torino

International collaborations

• Prof. S. Simoes - University of Coimbra, Portugal
• Prof. C. Viseras e C. Aguzzi - University of Granada, Spain
• Prof. H.A. Santos - University of Groningen, The Netherlands
• Prof. E. Csányi, S. Berko e M. Sucsz - University of Szged, Hungary
• Prof. A. Prina Mello - Trinity College Dublin, Ireland
• Prof. L. Black - Tufts University, Medford, MA, USA
• Prof D. Carugo - University College London, UK
• Prof. R. Donnelly e Dr A. Paredes - Queen’s University Belfast, UK

EUROPEAN PROJECTS

• European Commission H2020-NMBP-HUBS-2018, Project ID: 814607 “Safety testing in the life cycle of nanotechnology-enabled medical technologies for health” (2019-2023).
• European Commission HORIZON-CL4-2022-RESILIENCE-01-13 - Smart and multifunctional biomaterials for health innovations (RIA), Project ID:101092243 “smart and multiFunctional 3D printable prO-Regenerative biologiCal matrix modulating mEchanotRansduction as advancEd theraPy to treAt skIn chRonic wounds” “FORCEREPAIR” (2023-2026).

NATIONAL PROJECTS

• PNRR, M4 – Ecosistemi d’innovazione NODES – Nord Ovest Digitale E Sostenibile (2022-2025)
• PNRR, Project code PE00000003, “ON Foods - Research and innovation network on food and nutrition Sustainability, Safety and Security – Working ONFoods” (2022-2025).
• PNRR - One Health Basic and Translational Actions Addressing Unmet Needs on Emerging Infectious Diseases (INF-ACT) (2022-2025).

Expertise in the formulation development of: i) polymeric micro- and nanoparticles (Mini Spray Dryer 191 - Büchi; Nano Spray Dryer B-90 HP - Büchi), ii) micro- and nanofibers with random or aligned orientation (electrospinning equipment - STKIT-40 Linari Engineering - equipped with flat collector, rotating cylindrical collectors with different diameters and tool for the production of core-shell fibers; the equipment is placed in a temperature and humidity monitored cabined), iii) lipid nanosystems (Solid Lipid Nanoparticles, SLN, Nanostructured Lipid Carriers, NLC), iv) in situ gelling and/or mucoadhesive systems and v) polymeric matrices (films, porous matrices and multi-channel systems). Consolidated experience in the use of the following equipment for the characterization of the developed formulations: i) Particle Size and Zeta Potential Analyzer Litesizer 500 (Anton Paar) for size and surface potential determination of colloidal systems, ii) Malvern Mastersizer 3000E (Alfatest ) for particle size analysis of particle systems and liquid dispersions, iii) rotational rheometer MCR 102 (Anton Paar) for the study of the rheological behaviour of liquid and semi-solid systems (viscosity, viscoelasticity, thixotropy, rheological synergism), iv) tensiometer DyneMaster DY-300 (Kyowa) for the measurement of the surface and interfacial tension of solutions, v) FiveGo F3 conductivity meter (Mettler Toledo) for the evaluation of the conductivity of solutions, vi) TA.XT plus Texture Analyzer (Stable MicroSystems) for the study of the mechanical properties of solids (both in tension and in compression) and of the adhesive and bioadhesive properties of solids and semi-solids, vii) inclined plane apparatus for the evaluation of the mucoadhesive properties of liquids and semi-solids, viii) modified Franz cell for washability measurements on biological substrate ix) SurPASS 3 (Anton Paar) for the electrokinetic analysis of solid surfaces, x) Franz diffusion cells (with flat contact surface or with o-ring) (Permegear) for in vitro release studies and evaluation of hydration properties.

Expertise in statistical data processing and in the setting and analysis of experimental designs (DoE) using dedicated software (Statgraphics).

Continuous experience in the use of various cell lines a) human dermal fibroblasts, keratinocytes, endothelial cells (HUVEC), monocytes, tenocytes (Ten-1), osteoblasts (HOB), Schwann cells (SCs), human mesenchymal stem cells isolated from adipose tissue (hASC), HeLa cells, Caco-2 cells and Saos-2 cells for in vitro biopharmaceutical studies (cytotoxicity and cell proliferation studies). Equipment for in vitro studies on model cell lines: laminar flow cabinet (Space 2 Ergosafe, Pbi Int.), O2/CO2 incubator (mod. 5215, Pbi Int.), inverted optical microscope (mod. PQ 10.8, Pbi Int.), autoclave (Alfa Junior, Pbi Int.), centrifuge (mod. TC6, Sorvall). Acquired skills: preparation of culture media, seeding, expansion, freezing and thawing of cells, use of cell viability tests (MTT, Alamar Blue) and wound healing (use of inserts to evaluate the gap closure capacity), use of ELISA kit, fixation and staining of cellular samples for scanning electron microscopy and confocal microscopy investigations.